Guicheney P
INSERM U7, Department of Pharmacology, Hôpital Necker, Paris, France.
Methods Find Exp Clin Pharmacol. 1988 Apr;10(4):253-8.
Platelet serotonin (5-HT) content was determined from platelet rich plasma (PRP) of volunteers using high pressure liquid chromatography (HPLC) coupled with electrochemical detection following platelet lysis by perchloric acid. Several methodological and physiological aspects were studied. Blood collection with either EDTA or ACD-C anticoagulation gave identical results and blood storage at room temperature for at least two hours did not affect platelet 5-HT content, which allowed blood transfer to the laboratory. Deproteinized samples could be stored at either +4 degrees C for 48 hours or -80 degrees C for several weeks before HPLC determination. The partial degradation of 5-HT during storage was proportional to that of an internal standard at +4 degrees C or at -80 degrees C, but not at -20 degrees C. Comparison of platelet 5-HT content and serum 5-HT levels using the same HPLC method showed a good correlation although the absolute values differed. The mean platelet 5-HT content in healthy subjects aged 20-60 years was 0.364 + 0.093 nmol/10(+8) platelets (x + SD, n = 88). Men had slightly less platelet 5-HT than women and platelet 5-HT content was found to be lower in women over 60 years of age. This may come partly from hormonal suppression since platelet 5-HT contents were found to be positively correlated to plasma estrogen concentrations in a group of peri- and postmenopausal women.
使用高压液相色谱(HPLC)结合电化学检测法,从志愿者富含血小板的血浆(PRP)中测定血小板血清素(5-羟色胺,5-HT)含量,该检测在血小板经高氯酸裂解后进行。研究了几个方法学和生理学方面的问题。用乙二胺四乙酸(EDTA)或酸性枸橼酸盐葡萄糖溶液C(ACD-C)抗凝采集的血液结果相同,且血液在室温下储存至少两小时不影响血小板5-HT含量,这使得血液能够转运至实验室。在进行HPLC测定之前,去蛋白样品可以在4℃储存48小时或在-80℃储存数周。5-HT在储存期间的部分降解与内标在4℃或-80℃时的降解成比例,但在-20℃时不成比例。使用相同的HPLC方法比较血小板5-HT含量和血清5-HT水平,尽管绝对值不同,但显示出良好的相关性。20至60岁健康受试者的平均血小板5-HT含量为0.364 + 0.093 nmol/10⁸血小板(x + SD,n = 88)。男性的血小板5-HT含量略低于女性,且发现60岁以上女性的血小板5-HT含量较低。这可能部分源于激素抑制,因为在一组围绝经期和绝经后女性中发现血小板5-HT含量与血浆雌激素浓度呈正相关。